Detection of Staphylococcus aureus cell walls by enzyme-linked immunoassay using antibodies prepared from a semi-synthetic peptidoglycan precursor. 2012

Sandeep Sandhu, and James A Schouten, and Julie Thompson, and Mark Davis, and Timothy D H Bugg
Department of Chemistry, University of Warwick, Coventry, CV4 7AL, UK.

The peptidoglycan layer of Staphylococcus aureus contains a (Gly)(5) cross-link which is not found in other bacteria, and which could be used to develop a specific immunoassay for detection of S. aureus in MRSA infections. A semi-synthetic route was used to prepare the S. aureus peptidoglycan precursor UDPMurNAc-L-Ala-γ-D-Glu-L-Lys(Gly)(5)-D-Ala-D-Ala, which was covalently attached to carrier protein bovine serum albumin via the UDP nucleotide. Serum raised using this antigen showed specificity for chemically immobilised peptidoglycan monomer containing (Gly)(5), using an ELISA immunoassay. ELISA assays using 0.1 or 1.0 μg samples of cell walls prepared from two MRSA strains and one penicillin-sensitive S. aureus strain, and from three other bacteria, showed the highest response against cell walls containing (Gly)(5), with a particularly high response against cell walls from one MRSA strain. Competition assays to investigate antibody selectivity demonstrated that the antibody response could be most effectively antagonised using ligands containing (Gly)(5). These data demonstrate that it is possible to generate antibodies with high affinity and selectivity for the (Gly)(5) containing monomer in S. aureus peptidoglycan, that could be used to develop an immunoassay for S. aureus.

UI MeSH Term Description Entries
D007106 Immune Sera Serum that contains antibodies. It is obtained from an animal that has been immunized either by ANTIGEN injection or infection with microorganisms containing the antigen. Antisera,Immune Serums,Sera, Immune,Serums, Immune
D009842 Oligopeptides Peptides composed of between two and twelve amino acids. Oligopeptide
D010457 Peptidoglycan A structural polymer of the bacterial cell envelope consisting of sugars and amino acids which is responsible for both shape determination and cellular integrity under osmotic stress in virtually all bacteria. Murein,Pseudomurein
D002417 Cattle Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor. Beef Cow,Bos grunniens,Bos indicus,Bos indicus Cattle,Bos taurus,Cow,Cow, Domestic,Dairy Cow,Holstein Cow,Indicine Cattle,Taurine Cattle,Taurus Cattle,Yak,Zebu,Beef Cows,Bos indicus Cattles,Cattle, Bos indicus,Cattle, Indicine,Cattle, Taurine,Cattle, Taurus,Cattles, Bos indicus,Cattles, Indicine,Cattles, Taurine,Cattles, Taurus,Cow, Beef,Cow, Dairy,Cow, Holstein,Cows,Dairy Cows,Domestic Cow,Domestic Cows,Indicine Cattles,Taurine Cattles,Taurus Cattles,Yaks,Zebus
D002473 Cell Wall The outermost layer of a cell in most PLANTS; BACTERIA; FUNGI; and ALGAE. The cell wall is usually a rigid structure that lies external to the CELL MEMBRANE, and provides a protective barrier against physical or chemical agents. Cell Walls,Wall, Cell,Walls, Cell
D004797 Enzyme-Linked Immunosorbent Assay An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed. ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013211 Staphylococcus aureus Potentially pathogenic bacteria found in nasal membranes, skin, hair follicles, and perineum of warm-blooded animals. They may cause a wide range of infections and intoxications.

Related Publications

Sandeep Sandhu, and James A Schouten, and Julie Thompson, and Mark Davis, and Timothy D H Bugg
October 1985, Journal of clinical microbiology,
Sandeep Sandhu, and James A Schouten, and Julie Thompson, and Mark Davis, and Timothy D H Bugg
August 2000, Letters in applied microbiology,
Sandeep Sandhu, and James A Schouten, and Julie Thompson, and Mark Davis, and Timothy D H Bugg
January 1968, Journal of bacteriology,
Sandeep Sandhu, and James A Schouten, and Julie Thompson, and Mark Davis, and Timothy D H Bugg
July 1975, Zeitschrift fur Immunitatsforschung, experimentelle und klinische Immunologie,
Sandeep Sandhu, and James A Schouten, and Julie Thompson, and Mark Davis, and Timothy D H Bugg
February 1984, Acta pathologica, microbiologica, et immunologica Scandinavica. Section B, Microbiology,
Sandeep Sandhu, and James A Schouten, and Julie Thompson, and Mark Davis, and Timothy D H Bugg
July 1985, Journal of clinical pathology,
Sandeep Sandhu, and James A Schouten, and Julie Thompson, and Mark Davis, and Timothy D H Bugg
January 1985, Folia haematologica (Leipzig, Germany : 1928),
Sandeep Sandhu, and James A Schouten, and Julie Thompson, and Mark Davis, and Timothy D H Bugg
May 1983, Journal of clinical microbiology,
Sandeep Sandhu, and James A Schouten, and Julie Thompson, and Mark Davis, and Timothy D H Bugg
January 1989, Journal of immunoassay,
Sandeep Sandhu, and James A Schouten, and Julie Thompson, and Mark Davis, and Timothy D H Bugg
May 2016, Proceedings of the National Academy of Sciences of the United States of America,
Copied contents to your clipboard!